Immunohistochemical localization of galactosyltransferase in human fibroblasts and HeLa cells.
Anti-human galactosyltransferase (E.C. 2.4.1.22) antibodies were elicited in rabbits and purified on a galactosyltransferase-agarose column. Purified antibodies were used to localize galactosyltransferase in acetone-fixed HeLa cells and human lung fibroblasts. Both protein A-peroxidase developed with 3-amino 9-ethylcarbazole and swine anti-rabbit IgG-fluorescein isothiocyanate served to detect binding of anti-galactosyltransferase antibodies. In cells of confluent cultures, anti-galactosyltransferase staining appeared as a concise triangular structure in the juxtanuclear region with one angle oriented toward the bulk of the cytoplasm. The stained structure appeared as a dense cap on the nucleus in HeLa cells and as a more extended granular structure in fibroblasts. In cells of sparse cultures, specific anti-galactosyltransferase staining appeared in both HeLa cells and fibroblasts as a granular, extended structure, which was occasionally perinuclear. There was no evidence of cell surface localization of galactosyltransferase by light microscopy. The positively stained structures are interpreted to be part of the Golgi complex.
D000906:Antibodies; D000936:Antigen-Antibody Complex; D003902:Detergents / Q000494:pharmacology; D005347:Fibroblasts / Q000201:enzymology; D005455:Fluorescent Antibody Technique; D005700:Galactosyltransferases / Q000378:metabolism*; D006367:HeLa Cells / Q000201:enzymology; D006651:Histocytochemistry; D006801:Humans; D005779:Immunodiffusion; D017830:Octoxynol; D011092:Polyethylene Glycols / Q000494:pharmacology
D000906:Antibodies; D000936:Antigen-Antibody Complex; D003902:Detergents; D011092:Polyethylene Glycols; D017830:Octoxynol; D005700:Galactosyltransferases